Laser-Excited High-Sensitice Fluorescence Microscope System for Biological Cells using High-Speed Multi-Image Processing

使用高速多图像处理的生物细胞激光激发高灵敏度荧光显微镜系统

• 批准号: 59850067
• 负责人: INABA Humio
• 金额: $ 11.65万
• 依托单位: Tohoku University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Developmental Scientific Research
• 财政年份: 1984
• 资助国家: 日本
• 起止时间: 1984 至 1985
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-59850067/
• 关键词: Laser; Single Biologica Cell; Fluorescence Microscope; High-Speed Multi-Image Processing; 3-Dimensional Intensity Display; 螢光寿命測定

This project aimed to the new development of high-sensitive fluorescence microscope system, which enables us to measure quntitatively fluorescence or scattering intensity distribution in a single viological cell utilizing optoelectronic technique including lasers and multiple image processing methods.At first, We developes a laser-excited fluorescence microscope systen using an Ar laser for excitation, a SIT camera for sensitive detection and an image processor for digital processing of image signals. It was proved that this system can detect the fluorescence distribution from biological cell containing some organic dyes and analyze quantitatively the image signal from the SIT camara. Based on the results and understandings derived in the above study, we further improved the system employing an image intensifier and a new image processor to realize excellent high-sensitivity as well as better spatial and temporal resolutions. Thus, very weak fluorescence intensity distributions from se … More veral kinds of cells containing clinically interesting organic dyes were measured and displayed three-dimensionally with this system, which indicate accumulated portions of these dyes with in the cells. Temporal changes in fluorescence intensity distribution, which were mainly caused by the photochemical effect were also detected quatitatively in a short time interval less than 34 sec.Moreover, a laser-excited high-speed, time-resolved fluorescrnce mivroscope system was newly developed to investigate fluorescence decays from various dyes in biological cells in nanosecond time scale. Using this system incorporating an UV pulsed <N_2> laser, we performed characteristics measurement and comparison of photosensitizing hematoporphyrin species and pheophorbide a in cultured cancer cells and solutions for the first time and their different behaviors were discussed.In summary, we have succeeded in constructing a novel type of laser-excited high-speed fluorescnce microscope system for single biological cells using highspeed multi-image processing technique. The system and the results achieved by this Research Project should be useful and valuable in the fields of medicine, lifescience and viotechnology and their practical applications are expected in the near future. Less

本项目针对高灵敏度荧光显微镜系统的新发展，利用包括激光和多种图像处理方法在内的光电技术，定量测量单个生物细胞中的荧光或散射强度分布。首先，我们研制了一套以Ar激光器为激发光源、SIT摄像机为灵敏检测单元、图像处理单元为图像处理单元的激光激发荧光显微镜系统。实验证明，该系统可以检测含有有机染料的生物细胞的荧光分布，并对SIT摄像机的图像信号进行定量分析。基于以上研究的结果和认识，我们进一步改进了系统，采用了图像增强器和新的图像处理器，以实现优异的高灵敏度以及更好的空间和时间分辨率。因此，来自Se…的非常弱的荧光强度分布用该系统测量了多种含有临床上感兴趣的有机染料的细胞，并进行了三维显示，显示了这些染料在细胞中的累积部分。在不到34秒的短时间间隔内，定量检测到了主要由光化学效应引起的荧光强度分布的时间变化。此外，还新开发了一套激光激发的高速、时间分辨荧光显微镜系统，用于在纳秒时间尺度上研究各种染料在生物细胞中的荧光衰减。利用该系统结合紫外光脉冲氮气激光器，首次对培养的癌细胞和溶液中的光敏性血卟啉和脱镁叶绿酸a进行了特性测量和比较，并对它们的不同行为进行了讨论。综上所述，我们成功地利用高速多图像处理技术构建了一种新型的激光激发单个生物细胞的高速荧光显微镜系统。该系统和研究成果在医学、生命科学和病毒技术等领域具有一定的实用价值，并有望在不久的将来得到实际应用。较少

项目成果

日本レーザー医学会誌. 6-3. (1986)

日本激光医学会杂志 6-3 (1986)。

Japanese Journal of Optics. 14(4). (1985)

日本光学杂志。

応用物理. 53-2. (1984)

应用物理学。53-2。

Journal of Japan Society for Laser Medicine. 5(3). (1985)

日本激光医学会杂志。

日本レーザー医学会誌. 5-3. (1985)

日本激光医学会杂志 5-3 (1985)。